KASP primer combination suitable for lycopersicon esculentum DNA fingerprint database construction and application of KASP primer combination

A primer combination and primer set technology, which can be used in DNA/RNA fragmentation, recombinant DNA technology, determination/inspection of microorganisms, etc., and can solve problems such as rare and no special kits

Active Publication Date: 2019-12-24
北京通州国际种业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, there are few reports on the construction of tomato KASP-labeled nucleic acid fingerprint library, and there is no special kit for the construction of nucleic acid fingerprint library

Method used

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  • KASP primer combination suitable for lycopersicon esculentum DNA fingerprint database construction and application of KASP primer combination
  • KASP primer combination suitable for lycopersicon esculentum DNA fingerprint database construction and application of KASP primer combination
  • KASP primer combination suitable for lycopersicon esculentum DNA fingerprint database construction and application of KASP primer combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Embodiment 1, the screening of KASP primer combination

[0087] A large number of sequence analysis and functional analysis were performed on the tomato genome, and 48 SNP sites were screened (Table 2) (the physical coordinates refer to the tomato genome version 2.4, ftp: / / ftp.solgenomics.net / genomes / Solanum_lycopersicum / Heinz1706 / wgs / assembly / build_2.40 / ), and KASP primer design was carried out on it, and a large number of primers were screened among more than 1,000 pairs of primers designed to select primers with stable amplification effect and clear typing, and obtain the KASP primer combination constructed by the tomato DNA fingerprint library. Such as Table 3 (numbers in Table 3 correspond to those in Table 2) and figure 1 shown. Using the primers shown in Table 3 to carry out the results of the amplification detection of the materials shown in Table 1, see image 3 . Other primers (controls) in the screening process are used to carry out the results of amplifi...

Embodiment 2

[0095] Embodiment 2, construction and application of tomato DNA fingerprint library

[0096] 1. Construction of Tomato DNA Fingerprint Library

[0097] 1. Genomic DNA of 96 young leaves of tomato (Table 1) to be tested was extracted by CTAB method.

[0098] 2. Use 48 sets of primers in Table 3 to carry out PCR amplification on 96 tomato samples to be tested (48 sets of primers are in the same reaction system), and detect the genotypes of the tomato to be tested at 48 SNP sites. Fluorescent quantitative PCR instrument AB-Q6 Flex can be used for detection or the ArrayTape platform of Douglas Scientific company can be used for detection.

[0099]The reaction system (5 μL) when using the fluorescent quantitative PCR instrument AB-Q6 Flex (Thermo Fisher Scientific (China) Co., Ltd.) detection is: Genomic DNA (50ng / μL) 1.0 μL + primer mixture 0.7 μL (in the primer mixture The concentration of each primer in the set of primers in Example 1 is 100 pmol / L) + 2×KASP Mix (Aiji Analysis...

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Abstract

The invention discloses a KASP primer combination suitable for lycopersicon esculentum DNA fingerprint database construction and application of the KASP primer combination. Developed lycopersicon esculentum KASP core primers and a variety detection kit can be used for identifying the variety of to-be-detected lycopersicon esculentume, thus operation of distinguishing variety authenticity becomes accurate and convenient, the KASP primer combination is throughout a whole lycopersicon esculentum genome, comprises a unique amplification stable point, and has strong representativeness; and the kitis stable and reliable in detection result and rapid and easy to operate, and scale and standardized detection can be achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a set of KASP primer combinations suitable for constructing tomato DNA fingerprint libraries and applications thereof. Background technique [0002] Tomato (Lycopersicon esculentum L.) is one of the most important vegetable crops in the world with high nutritional value. Due to the unique edible, health care and auxiliary therapeutic effects of tomato and its products, it occupies an important position in the world's agricultural production and trade. According to statistics released by the World Processed Tomato Council (WPTC): In 2013, the global output of processed tomatoes was 33.012 million tons. Since tomato is a self-pollinated plant, in the process of hybrid seed production, if the artificial detasseling is not timely, the female parent will self-fertilize to produce false hybrids, and there will often be biological and mechanical hybrids, resulting in a decrease in the gene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 刘宝平王均帅焦珍珍邢海星张东峰马睿赵小翠尹增奇吴坤生
Owner 北京通州国际种业科技有限公司
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